Fasting-mimicking diet (FMD) but not water-only fasting promotes reversal of inflammation and IBD pathology

ABSTRACT

A method for treating autoimmune and/or inflammatory disease includes a step of identifying a subject exhibiting symptoms of autoimmune and/or inflammatory disease administering a fasting mimicking diet. A probiotic composition for gastrointestinal autoimmune and/or inflammatory disease Bacteroides acidifaciens, Bifidobacterium choerinum, and combinations thereof is also provided.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is the U.S. national phase of PCT Appln, No,PCT/US2019, 022488 filed Mar. 15, 2019 which claims the benefit of U.S.provisional patent application Ser. Nos. 62/643,296 filed Mar. 15, 2018;62/734,475 filed Sep. 21, 2018; and 62/735,147 filed Sep. 23, 2018, thedisclosures of which are hereby incorporated in their entirety byreference herein.

TECHNICAL FIELD

In at least one aspect, the present invention provides a treatment forgastrointestinal autoimmune/inflammatory disease by mediating positivechanges in the gut microbiome. The present invention combines thecomposition of a fasting-mimicking diet (FMD) with the regimen of a FMDplus refeeding cycles to enhance the cultivation and expansion ofbeneficial gut microbiota. In another aspect, the composition ofmicrobiota which can alone promote therapeutic effects againstgastrointestinal inflammatory diseases is described. Additionally, FMDplus refeeding cycles promoted the enhanced reversal of IBD pathology incomparison to 48 hr water-only fasting cycles.

BACKGROUND

Autoimmune diseases involve a miscommunication between innate andadaptive immunity and an imbalance between T lymphocytes populations,which play critical roles in the immuno-pathogenesis of manyautoimmune/chronic inflammatory diseases¹⁻³. It has been shown that bothhyperactive innate immune response (i.e. macrophages and dendriticcells), imbalance between autoreactive associated effector cells (i.e.CD4⁺Th1 and CD4⁺Th17) and anti-inflammatory associated regulatory Tcells (CD4⁺ Treg), and pro-inflammatory cytokine productions contributeto the pathogenesis of major gastrointestinal autoimmune/inflammatorydiseases including Crohn's disease (CD), ulcerative colitis (UC),irritable bowel syndrome, celiac disease, microscopic colitis(collagenous and lymphocytic colitis), and Bejcet disease³⁻⁸.

Microbiome and probiotic-based therapies for gastrointestinalautoimmune/inflammatory diseases currently under investigation primarilyinvolve approaches that include fecal microbiota transplantation (FMT),oral probiotic administration, or changes in diet⁹. While theseapproaches show promise, there is a need for fine-tuning these therapiesso they are suitable for treating patients affected by gastrointestinalautoimmune/inflammatory diseases clinically¹⁰.

SUMMARY

The present invention solves one or more problems of the prior art byproviding a method for treating or preventing autoimmune and/orinflammatory disease. The method includes a step of identifying asubject exhibiting symptoms of or at risk for autoimmune and/orinflammatory disease administering a periodic fasting mimicking diet.

In another embodiment, a probiotic composition for gastrointestinalautoimmune and/or inflammatory disease is provided. The probioticcomposition includes a bacterial component selected from the groupconsisting of Lactobacillaceae, Erysipelotrichaceae, andBifidobacteriaceae with the option of adding other beneficial microbialpopulations identified in the experiments set forth below, andcombinations thereof to be administered for the prevention and treatmentfor gastrointestinal autoimmune and/or inflammatory disease. Theprobiotic composition also includes an optional protective componentthat stabilizes the bacterial component.

The fasting-mimicking diet is also shown to enhance the reversal of IBDpathology in comparison to 48 hr water-only fasting cycles, as seen byimprovement in disease activity level, colon and small intestineregeneration, an increase in intestinal regeneration markers, and shiftsin the microbiome that promote the sustained expansion of probioticstrains.

In another embodiment, a dietary supplement for the prevention andtreatment of gastrointestinal autoimmune and/or inflammatory disease isprovided. The dietary supplement include pre-biotic ingredients and/orvegetables having such pre-biotic ingredients.

BRIEF DESCRIPTION OF THE DRAWINGS

-   FIG. 1A provides plot showing the most abundant taxa at the phylum    and family level.-   FIG. 1B provides a graphical representation of the top 3 microbiota    families (S24-7, Lactobacillaceae, Erysipelotrichaceae) and their    percent relative abundance mean at specific timepoints of diet    administration (C1=2 days after 4th DSS Cycle, F1=after completing    one FMD cycle and 2 Days after 4th DSS cycle, F2=after completing    four days of 2nd FMD cycle, C3=9 days after 4th DSS Cycle, F3=2 days    after 2nd FMD cycle and 9 days after 4th DSS cycle).-   FIG. 1C provides a table comparing population sizes of the top 8    microbiota families between the C3 and F3 time points. (C3=9 days    after 4th DSS Cycle, F3=2 days after 2nd FMD cycle and 9 days after    4th DSS cycle), as well as approximate fold change of the    populations at C3 vs. F3.-   FIG. 1D provides a Table outlining the most enriched gut microbiota    in the F3 group at the genus, species, and strain levels.-   FIG. 2A provides a visual representation of colon lengths of fecal    transplant (FT) recipients. Recipients were either given an FMT from    DSS control or DSS+FMD donors.-   FIG. 2B provides colon length quantification among Naive, DSS FT    Recipients, and DSS+FMD Recipients (One-way ANOVA, ***p<0.001).-   FIG. 2C provide a plot of Overall Disease Activity Index (DAI)    quantification after the 3rd cycle of DSS and through the fecal    transplant administration period.-   FIG. 2D provides a scatter plot of stool consistency after the 3rd    cycle of DSS and through the fecal transplant administration period.-   FIG. 3A provides a scatter plot of the Disease Activity Index (DAI)    scores of the Naive (n=15), DSS control diet, and DSS control diet    plus 2 cycles of FMD (DSS+FMD) starting after the third DSS cycle.-   FIG. 3B provides a plot of the Disease Activity Index (DAI) scores    of the Naive (n=15), DSS control diet, and DSS control diet plus 2    cycles of water-only fasting (DSS+WF) groups starting after the    third DSS cycle.-   FIG. 3C provides a plot of the stool consistency variable of the    Disease Activity Index (DAI) scores of the Naive, DSS control diet,    DSS control diet plus 2 cycles of FMD.-   FIG. 3D provides a plot of the stool consistency variable of the    Disease Activity Index (DAI) scores of the Naive, and DSS control    diet plus 2 cycles of water-only fasting groups starting after the    third DSS cycle.-   FIG. 3E provides a plot of the Hemoccult test variable of the    Disease Activity Index (DAI) scores of the Naive, DSS control diet,    and DSS control diet plus 2 cycles of FMD.-   FIG. 3F provides a plot of the Hemoccult test variable of the    Disease Activity Index (DAI) scores of the Naive, DSS control diet,    and DSS control diet plus 2 cycles of water-only fasting groups    starting after the third DSS cycle.-   FIG. 4A provides a visual representation of murine colon length from    Naïve, DSS control diet after 3 cycles (DSS 3 cycles), DSS control    diet after four cycles (DSS 4 cycles), DSS control diet after 4    cycles of DSS plus 2 cycles of FMD (DSS+FMD) and DSS control diet    plus 2 cycles of water-only fasting (DSS+WF) groups.-   FIG. 4B provides quantification of colon lengths of the Naive, DSS    control diet after 3 cycles, DSS control diet, DSS control diet plus    2 cycles of FMD, and DSS control diet plus 2 cycles of water-only    fasting. (One-way ANOVA, *p<0.05, **p<0.01, ***p<0.001).-   FIG. 5A provides a visual representation of murine small intestine    from Naïve, DSS control diet after 3 cycles (DSS 3 cycles), DSS    control diet after four cycles (DSS 4 cycles), DSS control diet    after 4 cycles of DSS plus 2 cycles of FMD (DSS+FMD) and DSS control    diet plus 2 cycles of water-only fasting (DSS+WF) groups.-   FIG. 5B provides quantification of small intestine lengths of the    Naïve, DSS control diet (DSS), DSS control diet plus 2 cycles of FMD    (DSS+FMD), and DSS control diet plus 2 cycles of water-only fasting    (DSS+WF). (One-way ANOVA, *p<0.05).-   FIG. 6A provides immunohistochemistry for BrdU⁺ cells and for    Lgr5+cells in proximal colonic crypts of murine colon ICC sections    in Naïve, DSS control diet (DSS), DSS control diet plus 2 cycles of    FMD (DSS+FMD) groups, and DSS control diet plus 2 cycles of    water-only fast (DSS+WF).-   FIG. 6B provides quantification of BrdU⁺ cells per proximal colonic    crypt in Naïve, DSS control diet (DSS), DSS control diet plus 2    cycles of FMD (DSS+FMD), and DSS control diet plus 2 cycles of    water-only fasting (DSS+WF) groups.-   FIG. 6C provides Quantification of Lgr5⁺ cells per proximal colonic    crypt in Naïve (n=8), DSS control diet (DSS), DSS control diet plus    2 cycles of FMD (DSS+FMD), and DSS control diet plus 2 cycles of    water-only fasting (DSS+WF) groups.-   FIG. 7A provides a plot showing microbiota shifts in the DSS+FMD    group 9 days after the 4th DSS cycle/two days after the 2nd FMD    cycle, and the DSS+WF group 9 days after the 4^(th) DSS cycle/four    days after the 2^(nd) water-only fast.-   FIG. 7B provides table summarizing the top 8 most abundant families    in fecal samples between the groups at these timepoints.-   FIG. 8A provides a plot of WBC count (10³/μl ) from patients with    low CRP (<1 mg/L; n=36) or high CRP (>1 mg/L) prior to dietary    intervention (a), at the end of an initial 5-day FMD cycle before    resuming normal food intake (b), and approximately 5 days after    completing 3 FMD cycles and refeeding (c).-   FIG. 8B provides a plot of circulating lymphocyte count (10³/μl )    from patients with low CRP (<1 mg/L; n=36) or high CRP (>1 mg/L)    prior to dietary intervention (a), at the end of an initial 5-day    FMD cycle before resuming normal food intake (b), and approximately    5 days after completing 3 FMD cycles and refeeding (c).-   FIG. 8C provides a plot of WBC counts (10³/μl ) in untreated, naive    mice or mice that received 4 cycles of DSS (a), on the last day of 1    cycle of a 4-day FMD between the 3^(rd) and last DSS cycles (b), and    two days after 4 DSS cycles and 2 FMD cycles (c).-   FIG. 8D provides a plot of Circulating lymphocyte counts (10³/μl )    in untreated, Naïve mice or mice that received 4 cycles of DSS (a),    on the last day of 1 cycle of a 4-day FMD between the 3^(rd) and    last DSS cycles (b), and two days after 4 DSS cycles and 2 FMD    cycles (c). Data are presented as mean±SEM.

DETAILED DESCRIPTION

Reference will now be made in detail to presently preferredcompositions, embodiments, and methods of the present invention whichconstitute the best modes of practicing the invention presently known tothe inventors. The Figures are not necessarily to scale. However, it isto be understood that the disclosed embodiments are merely exemplary ofthe invention that may be embodied in various and alternative forms.Therefore, specific details disclosed herein are not to be interpretedas limiting, but merely as a representative basis for any aspect of theinvention and/or as a representative basis for teaching one skilled inthe art to variously employ the present invention.

The term “comprising” is synonymous with “including,” “having,”“containing,” or “characterized by.” These terms are inclusive andopen-ended and do not exclude additional, unrecited elements or methodsteps.

The phrase “consisting of” excludes any element, step, or ingredient notspecified in the claim. When this phrase appears in a clause of the bodyof a claim, rather than immediately following the preamble, it limitsonly the element set forth in that clause; other elements are notexcluded from the claim as a whole.

The phrase “consisting essentially of” limits the scope of a claim tothe specified materials or steps, plus those that do not materiallyaffect the basic and novel characteristic(s) of the claimed subjectmatter.

It is also to be understood that this invention is not limited to thespecific embodiments and methods described below, as specific componentsand/or conditions may, of course, vary. Furthermore, the terminologyused herein is used only for the purpose of describing particularembodiments of the present invention and is not intended to be limitingin any way.

It must also be noted that, as used in the specification and theappended claims, the singular form “a,” “an,” and “the” comprise pluralreferents unless the context clearly indicates otherwise. For example,reference to a component in the singular is intended to comprise aplurality of components.

Throughout this application, where publications are referenced, thedisclosures of these publications in their entireties are herebyincorporated by reference into this application to more fully describethe state of the art to which this invention pertains.

Abbreviations:

“CRP” means c-reactive protein.

“DAI” means disease activity index.

“DSS” means dextran sulfate sodium.

“FMD” means fasting mimicking diet.

“FMT” means fecal microbiota transplant.

“FT” means fecal transplant.

“IBD” means Inflammatory bowel disease.

“WF” means water-only fasting.

The term “subject” refers to a human or animal, including all mammalssuch as primates (particularly higher primates), sheep, dog, rodents(e.g., mouse or rat), guinea pig, goat, pig, cat, rabbit, and cow.

The term “fasting mimicking and enhancing diet” means a diet that mimicsthe effects of fasting typically by providing a subject with at most50-75% of their normal caloric intake. However, if the fasting mimickingdiet composition is maintained, based on our current and previousfindings, partial disease prevention and treatment effects areanticipated even if 100% of the normal caloric intake is provided tosubjects. The term “fasting mimicking and enhancing diet” is sometimessimply referred to as a “fasting mimicking diet.” These diets includethose diets that have been referred to as fasting mimicking diets.Examples of useful fasting mimicking and enhancing diets and method formonitoring the effects of these diets on markers such as IGF-1 andIGFBP1 in the context of the present invention are set forth in U.S.patent application Ser. Nos. 14/273,946 filed May 9, 2014; 14/497,752filed Sep. 26, 2014; 12/910,508 filed Oct. 22, 2010; 13/643,673 filedOct. 26, 2012; 13/982,307 filed Jul. 29, 2013; 14/060,494 filed Oct. 22,2013; 14/178,953 filed Feb. 12, 2014; 14/320,996 filed Jul. 1, 2014;14/671,622 filed Mar. 27, 2015; the entire disclosure of these patentapplications is hereby incorporated by reference. The fasting mimickingdiet set forth in U.S. patent application Ser. Nos. 14/060,494 and14/178,953 are found to be particularly useful in the present invention.

The term “prebiotic” refers to food ingredients that promote the growthof beneficial microorganism in a subject's intestines. Typically, thefood ingredients are nondigestible food ingredients.

The term “probiotic” refers a substance or composition stimulates thegrowth of beneficial microorganisms, and in particular, growth ofbeneficial microorganism in a subject's intestines.

In an embodiment, a method for treating or preventing an autoimmuneand/or inflammatory disease is provided. Examples of these conditionsincludes Crohn's disease (CD), ulcerative colitis (UC), irritable bowelsyndrome, celiac disease, microscopic colitis (collagenous andlymphocytic colitis), and Bejcet disease. The method includes a step ofidentifying a subject exhibiting symptoms of autoimmune and/orinflammatory disease administering a fasting mimicking diet.Identification of these conditions can involve blood test, bariumx-rays, colonoscopy with or without biopsy, and the like. In arefinement, the fasting mimicking diet is administered for 2 or morecycles.

The FMD is administered to the subject for a first predetermined timeperiod. In some variations, the first predetermined time period is equalto or greater than, in increasing order of preference, 3, 5, 6, or 7days. In addition, the first predetermined time period is equal to orless than, in increasing order of preference, 20, 15, 10, or 8 days. Ina refinement, the first predetermined time period is 1, 2, 3, 4, 5, 6,7, 8, 9, or 10 days. In another refinement, the first predetermined timeperiod is 5 to 10 days. In some variations of the methods set forthherein, the fasting mimicking and enhancing diet is repeated atintervals. For example, the fasting mimicking and enhancing diet can beinitiated once a month for the duration of the subject's treatment whichcan be 3 months to a year or more (e.g., 1 to 5 years).

In some variations, the fasting mimicking diet for each of the methodsset forth herein provides at most, in increasing order of preference,50%, 40%, 30%, or 100% of the subject's normal caloric intake. In arefinement, the fasting mimicking diet provides at least, in increasingorder of preference, 5%, 10%, or 20% of the subject's normal caloricintake. The subject's normal caloric intake is the number of kcal thatthe subject consumes to maintain his/her weight. The subject's normalcaloric intake may be estimated by interviewing the subject or byconsideration of a subject's weight. As a rough guide, subject's normalcaloric intake is on average 2600 kcal/day for men and 1850 kcal/day forwomen. In certain instances, the fasting mimicking diet provides thesubject with from 700 to 1200 kcal/day. In a particularly usefulrefinement, the fasting mimicking diet provides a male subject ofaverage weight with at most 1100 kcal/day and a female subject ofaverage weight with at most 900 kcal/day. In some refinements, thefasting mimicking diet provides at most, in increasing order ofpreference, 1500 kcal/day, 1400 kcal/day, 1300 kcal/day, 1200 kcal/day,1100 kcal/day, 1000 kcal/day, 900 kcal/day, 800 kcal/day, 700 kcal/day,600 kcal/day, 500 kcal/day, or 2500 kcal/day. In some furtherrefinements, the fasting mimicking diet provides at least, in increasingorder of preference, 0 kcal/day, 10 kcal/day, 100 kcal/day, 200kcal/day, 300 kcal/day, 400 kcal/day, or 500 kcal/day.

In certain variations, the fasting mimicking and enhancing diet providesfrom 4.5 to 7 kilocalories per pound of subject for a first day (day 1)and then 3 to 5 kilocalories per pound of subject per day for the secondto the final day. Day 1 ingredients are a blend of beets, carrots,collard, spinach, kale, mushroom, tomato, extra virgin olive oil,essential fatty acids, chicken broth, and veggie broth, while Day 2-4 isrestricted to chicken broth, veggie broth, and glycerol. The compositionof the Day 1 FMD is approximately 25% of a blend of the vegetableslisted above, 60% extra virgin olive oil and essential fatty acids, and10% of the broth mix. After a cycle of the fasting mimicking andenhancing diet, a second diet is administered to the subject for asecond predetermined time period. In a refinement, the second dietprovides an overall calorie consumption that is within 20 percent of asubject's normal calorie consumption for 10 to 26 days (e.g.,immediately) following the fasting mimicking and enhancing diet.

Notably, the relative protein, carbohydrate and fat content of the FMDcan be varied to achieve similar results based on total calorie intakeand length. This can be assessed based on IGF-1, IGFBP1, glucose andketone bodies. For example, a diet lasting 1 week and containing arelatively high level of protein and carbohydrates but providing 10-20%of the normal calorie intake can have similar fasting mimickingproperties to a diet lasting 5 days but containing low levels of carbsand proteins and providing 50% of normal calories. In other words, thisapplication describes fasting mimicking properties that providebeneficial effects which are variable based on relative macronutrientcomposition, diet length and calorie intake. The formulations providedabove are examples based on the discoveries made here and previouslywhich do not include all of the fasting mimicking compositions andmethods which are obvious to one skilled in the art to achieve therequired IGF-1, IGFBP1, ketone bodies, and glucose levels achieved bythe FMDs described here.

Additional examples of useful fasting mimicking and enhancing diets inthe context of the present invention are set forth in U.S. patentapplication Ser. Nos. 14/273,946 filed May 9, 2014; 14/497,752 filedSep. 26, 2014; 12/910,508 filed Oct. 22, 2010; 13/643,673 filed Oct. 26,2012; 13/982,307 filed Jul. 29, 2013; 14/060,494 filed Oct. 22, 2013;14/178,953 filed Feb. 12, 2014; 14/320,996 filed Jul. 1, 2014;14/671,622 filed Mar. 27, 2015; the entire disclosure of these patentapplications is hereby incorporated by reference. The fasting mimickingdiet set forth in U.S. patent application Ser. Nos. 14/060,494 and14/178,953 are found to be particularly useful in the present invention.Additional examples of FMD diets are found in U.S. patent applicationSer. No. 15/148,251 and WIPO Pub. No. WO2011/050,302 and WIPO Pub. No.WO2011/050,302; the entire disclosures of which are hereby incorporatedby reference and attached as exhibits A-H. Details of a particularlyuseful diet, the PROLON diet are found in U.S. patent application Ser.No. 15/432,803 filed Feb. 14; the entire disclosure of which is herebyincorporated by reference.

In a variation, it was found that two cycles of FMD, followed by twodays of refeeding (with the subjects normal diet), increased acombination of gut microbiota at the family level (Lactobacillaceae,Erysipelotrichaceae, Verrucomicrobiaceae, and Bifidobacteriaceae) whiledecreasing others (S24-7, Turicibacteraceae, Lachnospiraceae, andRuminococcaceae) when compared to the gut microbiota composition inducedby a control diet (FIG. 1A-C). Within the families Lactobacillaceae,Erysipelotrichaceae, and Bifidobacteriaceae, the genera Lactobacillus,Allobaculum, and Bifidobacterium were enriched, respectively, after twocycles of FMD followed by two days of refeeding. At the microbial strainlevel, Bacteroides acidifaciens and Bifidobacterium choerinum wereuniquely enriched in this group (FIG. 1D), both of which have been notedto prevent obesity and display probiotic activity to treat intestinaldisorders, respectively^(11,12).

In another embodiment, a probiotic composition for gastrointestinalautoimmune and/or inflammatory disease is provided. The probioticcomposition includes a bacterial component selected from the microbialstrains consisting of Bacteroides acidifaciens and Bifidobacteriumchoerinum, and combinations thereof to be administered for theprevention and treatment for gastrointestinal autoimmune and/orinflammatory disease. The probiotic composition can also include anoptional protective component that stabilizes the bacterial component.In a variation, the probiotic composition further includes gutmicrobiota strains isolated from the genera Lactobacillus, Allobaculum,and Bifidobacterium. In another variation, the bacterial componentincludes a combination of Bacteroides acidifaciens and Bifidobacteriumchoerinum such that the Bifidobacterium choerinum enhances the probioticactivity of Bacteroides acidifaciens and/or Bacteroides acidifaciensenhances the probiotic activity of Bifidobacterium choerinum. Theprobiotic composition can be administered to a patient in an effectiveamount to relieve symptoms of gastrointestinal autoimmune and/orinflammatory disease with or without an FMD.

In a variation, the probiotic composition of the invention includes atleast 10³ CFU/g of each of Bacteroides acidifaciens, Bifidobacteriumchoerinum, and optionally the strains isolated from the generaLactobacillus, Allobaculum, and Bifidobacterium in a capsule. Thebacterial species therefore are present in the dose form as livebacteria, whether in dried, lyophilized, or sporulated form. Forexample, the dose form can be a capsule containing the bacterialcomponents in a dried form, blended with a suitable carrier. A typicalprobiotic composition will include from about 10³ to about 10¹⁴ CFU/g ofeach bacterial component. In a refinement, the probiotic compositionincludes 10⁵-10¹² CFU/g of each bacterial component. In anotherrefinement, the probiotic composition includes 10⁹-10¹³ colony formingunits/g of each bacterial component. In another refinement, theprobiotic composition includes 10⁵-10⁷ CFU/g of each bacterialcomponent. In still another refinement, the probiotic compositionincludes 10⁸-10⁹ CFU/g.

As set forth above, examples of protective components and methods forstabilizing probiotics such as the probiotic composition of the presentinvention is found in Different Methods of Probiotics Stabilization,Kamila Goderska, Oct. 3, 2012 DOI: 10.5772/50313; the entire disclosureof which is hereby incorporated by reference. In a variation, theprotect component can be a protective carrier and/or protective coatingand/or a protective encapsulant. In a refinement, the protective carrieris a liquid at 25° C. (i.e., liquid carrier) with the bacterialcomponent dispersed therein. Examples of such liquids includehydroxylated hydrocarbon carriers such as polyol (e.g., glycerol). In avariation, these liquids interact with the bacterial component therebyforming a modified bacterial component. Such interactions can includechemical bonding (e.g., hydrogen bonding and Van der Waals bonds).Moreover, the liquid carrier can adsorb (e.g., including adsorption) tothe bacteria cell wall and/or cell membrane.

In a variation, the protective component includes a stabilization agent(e.g., a compound) that interacts (e.g., chemical bonding, adsorbing,and adsorption). with or encapsulates the bacterial component. Examplesof such compounds include, but are not limited to, adonitol, betaine,carbohydrates (e.g., sugars), proteins, amino acids, mixtures of sugarand protein (e.g., protectans), gums and skim milk. In this regard,protein can form relatively stable intracellular glasses. In arefinement, the stabilization agent is dissolved or dispersed in a roomtemperature liquid. Finally, the liquid carrier, the protectivecompound, or the protective coating can be contained in the interior ofthe bacteria (e.g., by diffusion or ingestion).

In another variation the protective component is an protect encapsulant.The bacterial component can be encapsulated by spray drying, fluidizedbed drying and vacuum drying. In a refinement, the protective componentincludes an encapsulating agent selected from the group consisting ofmaltodextrins, skim milk, reconstituted skim milk, casein, soybeanprotein, trehalose, maltodextrin, and combinations thereof. In a furtherrefinement, the protective component includes a prebiotic such asinulin, oligofructose, and oligofructose-enriched inulin

The bacterial compositions of the invention may additionally andoptionally include an optional additive selected from any suitableadjuvants, excipients, additives, additional carriers, additionaltherapeutic agents, bioavailability enhancers, side-effect suppressingcomponents, diluents, buffers, flavoring agents, binders, preservativesor other ingredients and combinations thereof that do not preclude theefficacy of the composition. In a refinement, the bacterial compositionsare present in an amount from about 50% to about 90% by weight of thecomposition and 10% to 50 weight percent of an optional additive.

In a variation, the probiotic composition can be administered along themethod for preventing or treating gastrointestinal autoimmune and/orinflammatory disease set forth above. In this regard, the probioticcomposition can be administered on each day or any subset of days thatthe FMD diet is administered. In a refinement, the probiotic compositioncan be administered on any day on which the FMD diet is notadministered.

In another embodiment, a dietary supplement for the prevention andtreatment of gastrointestinal autoimmune and/or inflammatory disease isprovided. The dietary supplement include pre-biotic ingredients orvegetable having such pre-biotic ingredients. Table 1 provides theprebiotic components found in several vegetables. In a refinement, thedietary supplement includes a vegetable mixture of beets, carrots,collard, spinach, kale, mushroom, tomato, and optionally nettle leaf. Ina refinement, the vegetable mixture can also include extra virgin oliveoil, essential fatty acids, and/or vegetable broth. Typically, thevegetable mixture incudes 5 to 20 weight percent beets, 5 to 20 weightpercent carrots, 5 to 20 weight percent collard, 5 to 20 weight percentspinach, 5 to 20 weight percent kale, 5 to 20 weight percent mushroom, 5to 20 weight percent tomato, and 0 to 20 weight percent nettle leaf. Ina refinement, the , the vegetable mixture incudes 8 to 15 weight percentbeets, 8 to 15 weight percent carrots, 8 to 15 weight percent collard, 8to 15 weight percent spinach, 8 to 15 weight percent kale, 8 to 15weight percent mushroom, 8 to 15 weight percent tomato, and 5 to 15weight percent nettle leaf.

In a refinement, the vegetable mixture is a powdered vegetable mixturethat is formed by desiccating (e.g., drying) the vegetable componentsand grinding or mechanically manipulating into a powder to form a driedvegetable powder. In a variation, the order of these steps can beinterchanged (e.g., grinding and then drying). It should be appreciatedthat the drying should be performed without heating to preserve vitamincontent. Examples of desiccating the vegetable mixture includesfreeze-drying, convection drying, spray drying, and the like. In arefinement, the dried vegetable powder has a plurality of particles witha size (e.g., diameter or largest spatial dimension) in the range of 1to 10 microns. In a further refinement, the dried vegetable powder hasan average size (e.g., diameter or largest spatial dimension) in therange of 1 to 10 microns. Of course, particles of this size range havealtered material properties (e.g., absorption properties, ingestability,and the like) as compared to the naturally occurring vegetables.Consistent with the ranges set forth above, the dried vegetable powderincudes 5 to 20 weight percent of powder formed from beets, 5 to 20weight percent of powder formed from carrots, 5 to 20 weight percentcollard, 5 to 20 weight percent of powder formed from spinach, 5 to 20weight percent of powder formed from kale, 5 to 20 weight percentmushroom, 5 to 20 weight percent of powder formed from tomato, and 0 to20 weight percent of powder formed from nettle leaf. In a refinement,the , the vegetable mixture incudes 8 to 15 weight percent of powderformed from beets, 8 to 15 weight percent of powder formed from carrots,8 to 15 weight percent of powder formed from collard, 8 to 15 weightpercent of powder formed from spinach, 8 to 15 weight percent of powderformed from kale, 8 to 15 weight percent of powder formed from mushroom,8 to 15 weight percent of powder formed from tomato, and 5 to 15 weightpercent of powder formed from nettle leaf. A useful batch of the dietsupplement will provide a dry powder equivalent of 5 servings ofvegetables (powder equivalent to 375 grams of raw vegetables, total).Typically, the dietary supplement is combined with water and optionallyheated to be consumed as a broth.

In a refinement, the dietary supplement is administered to a subjectseveral times a week. The subject may have been identified as having IBDand/or gastrointestinal autoimmune and/or inflammatory disease. In afurther refinement, the dietary supplement can be administered 1, 2, 3,4, 5, 6, or 7 days a week. In still a further refinement, the dietarysupplement can be taken twice as part of a week as part of a protocolagainst autoimmunities (e.g., once at day 1 and once in day 4 of anydiet such as the FMD set forth above). In this regard, the dietarysupplement can be combined with the probiotic composition and/or themethod for preventing or treating gastrointestinal autoimmune and/orinflammatory disease set forth above.

TABLE 1 Prebiotic components of several vegetables. Vegetable PrebioticIngredients Beet Root Cellulose, pectin/Pectic polysaccharides CarrotRoot Arabinogalactans Collard Leaf Soluble fibers, glycosylates KaleLeaf Soluble fibers, glucosinolates Nettle Leaf Terpenoids, carotenoids,chlorophyll, vitamins, tannins, carbohydrates, sterols, polysaccharides,isolectins, polyphenols, oleanol acid, sterols and steryl glycosidesSpinach Leaf Hydroxycinnamic acids: (E)-ferulic acid and (E)-p-coumaricacid; pectic polysaccharides Tomato Fruit Arabinogalactans,oligofructose Maitake Mycelium Polysaccharides: starch, naturaloligofructoses, fructo-oligosacharides (FOS), lactulose, andgalactomannan

The following examples illustrate the various embodiments of the presentinvention. Those skilled in the art will recognize many variations thatare within the spirit of the present invention and scope of the claims.

An FMD was administered in mice displaying signs for gastrointestinalautoimmune and/or inflammatory disease to further confirm that thiscombination of FMD gut microbiota executes positive changes onIBD-associated phenotypes (see, FIGS. 1 and 2).

The experimental mouse FMD is based on a nutritional screen thatidentified ingredients which allow high nourishment during periods oflow-calorie consumption, modeled after the same ingredients used in thehuman version of the FMD. The FMD diet consists of two differentcomponents designated as day 1 diet and day 2-4 diet that were fed inthis order respectively. Day 1 ingredients are a blend of beets,carrots, collard, spinach, kale, mushroom, tomato, extra virgin oliveoil, essential fatty acids, chicken broth, and veggie broth, while Day2-4 is restricted to chicken broth, veggie broth, and glycerol. Day 1diet contains 7.87 kJ/g, the day 2-4 diet is identical on all feedingdays and contains 1.51 kJ/g. Day 1 and day 2-4 diets were supplied tothe FMD cohort with the average intake of the ad lib control group (−4g) every two weeks. On average, mice consumed 11.07 kJ (plant-basedprotein 0.75 kJ, carbohydrate 5.32 kJ, fat 5 kJ) on each day of the FMDregimen. Mice consumed all the supplied food on each day of the FMDregimen and showed no signs of food aversion.

FIG. 1 shows that two, 4-day FMD cycles induces changes in gutmicrobiota at the phylum and family level. FIG. 1A provides plotsshowing the most abundant taxa at the phylum and family level while FIG.1B provide a bar chart of the top 3 microbiota families (S24-7,Lactobacillaceae, Erysipelotrichaceae) and their percent relativeabundance mean at specific timepoints of diet administration (C1=2 daysafter 4th DSS Cycle, F1=after completing one FMD cycle and 2 Days after4th DSS cycle, F2=after completing four days of 2nd FMD cycle, C3=9 daysafter 4th DSS Cycle, F3=2 days after 2nd FMD cycle and 9 days after 4thDSS cycle). (C) Table comparing population sizes of the top 8 microbiotafamilies between the C3 and F3 time points. (C3=9 days after 4th DSSCycle, F3=2 days after 2nd FMD cycle and 9 days after 4th DSS cycle), aswell as approximate fold change of the populations at C3 vs. F3. FIG. 1Dprovides a table outlining the most enriched gut microbiota in the F3group at the genus, species, and strain levels.

FIGS. 2A-C provides results of a fecal microbiota transplant (FMT)derived from mice fed with a human FMD diet induces changes in mousecolon length and improved disease outcome. FIG. 2A provide a visualrepresentation (i.e., a photograph) showing colon lengths of fecaltransplant (FT) recipients. Recipients were either given an FMT from DSScontrol or DSS+FMD donors. DSS causes a (DSS)-induced colitis model thatis commonly used to study IBD in mice.^(13,14) DSS is a sulfatedpolysaccharide that is especially toxic to the colonic epithelium. FIG.2B provides colon length quantification among Naive, DSS FT Recipients,and DSS+FMD Recipients (One-way ANOVA, ***p<0.001). FIGS. 2C and 2Dprovide overall Disease Activity Index (DAI) quantification and stoolconsistency after the 3rd cycle of DSS and through the fecal transplantadministration period.

FIGS. 3A-C provide experimental comparison of disease activity in micetreated with 4 DSS cycles, and either 2 FMD cycles or 2 water-onlyfasting cycles. Mice treated with FMD cycles have improvement in diseaseactivity, firmer stools (lower score for stool consistency), and lesspresence of blood in stools (lower score for Hemoccult) in comparison tomice treated with 48 hr water-only fasting cycles FIGS. 3A and 3Bprovide plots of the Disease Activity Index (DAI) scores of the Naive(n=15), DSS control diet, and DSS control diet plus 2 cycles of FMD(DSS+FMD) or DSS control diet plus 2 cycles of water-only fasting(DSS+WF) groups starting after the third DSS cycle. FIGS. 3C and 3Dprovide plots of the stool consistency variable of the Disease ActivityIndex (DAI) scores of the Naïve , DSS control diet, DSS control dietplus 2 cycles of FMD, and DSS control diet plus 2 cycles of water-onlyfasting groups starting after the third DSS cycle. FIGS. 3E and 3Fprovide the Hemoccult test variable of the Disease Activity Index (DAI)scores of the Naïve, DSS control diet, DSS control diet plus 2 cycles ofFMD, and DSS control diet plus 2 cycles of water-only fasting groupsstarting after the third DSS cycle. (Two-way ANOVA, *p<0.05, **p<0.01,***p<0.001, ****p<0.0001).

FIGS. 4A-B shows that regeneration of colon is enhanced in the DSS+FMDgroup compared to the DSS+WF group after 4 cycles of DSS. FIG. 4Aprovides a visual representation of murine colon length from Naïve, DSScontrol diet after 3 cycles (DSS 3 cycles), DSS control diet after fourcycles (DSS 4 cycles), DSS control diet after 4 cycles of DSS plus 2cycles of FMD (DSS+FMD) and DSS control diet plus 2 cycles of water-onlyfasting (DSS+WF) groups. FIG. 4B provides quantification of colonlengths of the Naïve, DSS control diet after 3 cycles, DSS control diet,DSS control diet plus 2 cycles of FMD, and DSS control diet plus 2cycles of water-only fasting. (One-way ANOVA, *p<0.05, **p<0.01,***p<0.001).

FIGS. 5A and 5B provide experimental results showing regeneration ofsmall intestine is enhanced in the DSS+FMD group with no change in theDSS+WF group after 4 cycles of DSS. FIG. 5A provides a visualrepresentation of murine small intestine from Naive, DSS control dietafter 3 cycles (DSS 3 cycles), DSS control diet after four cycles (DSS 4cycles), DSS control diet after 4 cycles of DSS plus 2 cycles of FMD(DSS+FMD) and DSS control diet plus 2 cycles of water-only fasting(DSS+WF) groups. FIG. 5B provides quantification of small intestinelengths of the Naïve, DSS control diet (DSS), DSS control diet plus 2cycles of FMD (DSS+FMD), and DSS control diet plus 2 cycles ofwater-only fasting (DSS+WF). (One-way ANOVA, *p<0.05).

FIG. 6A provide experimental results showing that markers forregeneration in the colon (BrdU and Lgr5) are increased in the DSS+FMDand DSS+WF group, with the DSS+FMD group having a greater increase inLgr5 in colonic crypts. FIG. 6A shows immunohistochemistry for BrdU⁺cells and for Lgr5+cells in proximal colonic crypts of murine colon ICCsections in Naïve, DSS control diet (DSS), DSS control diet plus 2cycles of FMD (DSS+FMD) groups, and DSS control diet plus 2 cycles ofwater-only fast (DSS+WF). FIG. 6B provides quantification of BrdU⁺ cellsper proximal colonic crypt in Naïve, DSS control diet (DSS), DSS controldiet plus 2 cycles of FMD (DSS+FMD), and DSS control diet plus 2 cyclesof water-only fasting (DSS+WF) groups. FIG. 6C provides quantificationof Lgr5⁺ cells per proximal colonic crypt in Naïve (n=8), DSS controldiet (DSS), DSS control diet plus 2 cycles of FMD (DSS+FMD), and DSScontrol diet plus 2 cycles of water-only fasting (DSS+WF) groups.(one-way ANOVA, *p <0.05, **p <0.01, ***p <0.001, and ****p<0.0001).

FIG. 7A provides a plot showing microbiota shifts in the DSS+FMD group 9days after the 4th DSS cycle/two days after the 2nd FMD cycle, and theDSS+WF group 9 days after the 4^(th) DSS cycle/four days after the2^(nd) water-only fast. FIG. 7B provides a table summarizing the top 8most abundant families in fecal samples between the groups at thesetimepoints. Lactobacilaceae is reduced in the DSS+WF group compared tothe DSS+FMD group (25.8+3.97% vs. 45.2+4.2%), as well as inErysipelotrichaceae (0.286+0.184% vs. 10.5+5.71%), with no detectablepresence of Bifidobacteriaceae. Paraprevotellaceae is present in theDSS+WF group but not in the DSS+FMD group (6.13+0.148%).

FIG. 8A-D provide plots showing that white blood cell (WBC) andlymphocyte counts in humans and mice with systemic inflammation improvesafter treatment with FMD cycles. FIG. 8A provides a plot of WBC count(10³/μl ) from patients with low CRP (<1 mg/L; n=36) or high CRP (>1mg/L) prior to dietary intervention (a), at the end of an initial 5-dayFMD cycle before resuming normal food intake (b), and approximately 5days after completing 3 FMD cycles and refeeding (c). FIG. 8A provides aplot of circulating lymphocyte count (10³/μl ) from patients with lowCRP (<1 mg/L; n=36) or high CRP (>1 mg/L) prior to dietary intervention(a), at the end of an initial 5-day FMD cycle before resuming normalfood intake (b), and approximately 5 days after completing 3 FMD cyclesand refeeding (c). FIG. 8C provides a plot of WBC counts (10³/μl ) inuntreated, naïve mice or mice that received 4 cycles of DSS (a), on thelast day of 1 cycle of a 4-day FMD between the 3^(rd) and last DSScycles (b), and two days after 4 DSS cycles and 2 FMD cycles (c). FIG.8D provides a plot of Circulating lymphocyte counts (10³/μl ) inuntreated, naïve mice or mice that received 4 cycles of DSS (a), on thelast day of 1 cycle of a 4-day FMD between the 3^(rd) and last DSScycles (b), and two days after 4 DSS cycles and 2 FMD cycles (c). Dataare presented as mean±SEM. (one-way ANOVA, *p <0.05, **p <0.01, and ***p<0.001).

Details of an Exemplary Prolon™ Fasting Mimicking Diet

In an embodiment of the present invention, a diet package foradministering a fasting mimicking diet is provides. The fastingmimicking diet package provides daily meal portions for a predeterminednumber of days. Typically, the predetermined number of days is from 1 to10 days (i.e., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 days). In a particularlyuseful variation, the predetermined number of days is 5 or 6 days. Insome variations, the fasting mimicking diets set forth herein provide asubject at most, in increasing order of preference, 75%, 50%, 40%, 30%,or 10% of the subject's normal caloric intake or the daily recommendedcaloric intake for a subject. In a refinement, the fasting mimickingdiet provides at least, in increasing order of preference, 5%, 10%, or20% of the subject's normal caloric intake or the daily recommendedcaloric intake for a subject. However, if the fasting mimicking dietcomposition is maintained, based on our current and previous findings,partial disease prevention and treatment effects are anticipated even if100% of the normal caloric intake is provided to subjects. The subject'snormal caloric intake is the number of kcal that the subject consumes tomaintain his/her weight. The subject's normal caloric intake may beestimated by interviewing the subject or by consideration of a subject'sweight. As a rough guide, subject's normal caloric intake is on average2600 kcal/day for men and 1850 kcal/day for women. In certain instances,the fasting mimicking diet provides the subject with 700 to 1200kcal/day. In a particularly useful refinement, the fasting mimickingdiet provides a male subject of average weight with about 1100 kcal/dayand a female subject of average weight with 900 kcal/day. In somevariation, the diet from the diet package is administered on consecutivedays. In another variation, the daily meal portions provided for onlyone day a week for at least a month.

In one embodiment, the fasting mimicking diet package providing dailymeal portions for a predetermined number of days as set forth above. Thefasting mimicking diet package includes a kale cracker composition, afirst vegetable broth composition, a mushroom soup composition, a tomatosoup composition, a quinoa-containing minestrone soup composition, abean-containing minestrone soup composition, and a pumpkin soupcomposition. Characteristically, the daily meal portions are packagedinto meal servings or into a total daily serving to be divided intomeals. In a refinement, the fasting mimicking diet package furtherincludes a nut-containing nutrition bar, a cocoa-containing nutritionbar, a first olive-containing composition, a first vegetable brothcomposition, a tea composition that includes spearmint, a energy drinkcomposition, a micro-nutritional composition, and a algal oilcomposition. In a further refinement, the fasting mimicking diet packagefurther includes a second olive-containing composition, a secondvegetable broth composition, a tea composition that includes spearmintand lemon, and a tea composition that includes hibiscus.

In a variation of the embodiments set forth above, the fasting mimickingdiet package includes daily meal portions that provide less than 40grams of sugar for day 1, less than 30 grams of sugar for days 2 to 5and any remaining days, less than 28 grams of protein for day 1, lessthan 18 grams of protein for days 2 to 5 and any remaining days, 20-30grams of monounsaturated fats or more to reach the desired caloricintake (i.e., a predetermined caloric intake) for day 1, 6-10 grams ofpolyunsaturated fats or more to reach the desired caloric intake for day1, 2-12 grams of saturated fats or more to reach the desired caloricintake for day 1, 10-15 grams of monounsaturated fats or more to reachthe desired caloric intake for days 2 to 5 and any remaining days, 3-5grams of polyunsaturated fats or more to reach the desired caloricintake for days 2 to 5 and any remaining days, 1-6 grams of saturatedfats or more to reach the desired caloric intake for days 2 to 5, or anyremaining days, and a micronutrient composition on each day and anyremaining days.

In another variation of the embodiments set forth above, the fastingmimicking diet package includes daily meal portions 8-10 kcal perkilogram body weight for each diet day. In this variation, the fastingmimicking diet provides less than 30 grams of sugar for each diet day,less than 18 grams of protein for each diet day, 9-15 grams ofmonounsaturated fats or more to reach the desired caloric intake foreach diet day, and 2.5-4.5 grams of polyunsaturated fats or more toreach the desired caloric intake for each diet day and 1-5.5 grams ofsaturated fats or more to reach the desired caloric intake for each dietday. Higher levels of the fats listed above can be provided for higherFMD formulation providing up to 100% of the normal caloric intake tosubjects.

In still another variation of the embodiments set forth above, thefasting mimicking diet package includes daily meal portions that provide5-8 kcal per kilogram body weight for each diet day. In this variation,the fasting mimicking diet provides less than 20 grams of sugar for eachdiet day, less than 12 grams of protein for each diet day, and 6.5-10grams of monounsaturated fats or more to reach the desired caloricintake for each diet day, 2.5-4.5 grams of polyunsaturated fats or moreto reach the desired caloric intake for each diet day and 1.5-4 grams ofsaturated fats or more to reach the desired caloric intake for each dietday.

In still another variation of the embodiments set forth above, thefasting mimicking diet package includes daily meal servings that provide0-3 kcal per kilogram body weight for each diet day. In this variation,the fasting mimicking diet provides less than 5 grams of sugar for eachdiet day, less than 3 grams of protein for each diet day, and less than2.5 grams of monounsaturated fats for each diet day, less than 1 gramsof polyunsaturated fats for each diet day and less than 1 grams ofsaturated fats for each diet day.

In an embodiment, the nutritional requirements for the fasting mimickingdiet set forth above can be realized by a diet package with certainspecific meal components. In one variation, the fasting mimicking dietpackage provides daily meal portions for a predetermined number of daysare set forth above (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 days). Thefasting mimicking diet package includes a kale cracker composition , afirst vegetable broth composition, a mushroom soup composition, a tomatosoup composition, a quinoa-containing minestrone soup composition, abean-containing minestrone soup composition, and a pumpkin soupcomposition, Characteristically, the daily meal portions are packagedinto meal servings or into a total daily serving to be divided intomeals. In a refinement, the fasting mimicking diet package furtherincludes a nut-containing nutrition bar, a cocoa-containing nutritionbar, a first olive-containing composition, a first vegetable brothcomposition, a tea composition that includes spearmint, a energy drinkcomposition, a micronutritional composition, and a algal oilcomposition. In a further refinement, the fasting mimicking diet packagefurther includes a second olive-containing composition, a secondvegetable broth composition, a tea composition that includes spearmintand lemon, and a tea composition that includes hibiscus. It should beappreciated that each of the soup, broth, tea and energy compositionsset forth herein are designed to have added water when consumed.

In another variation of a fasting mimicking diet package, diet packageincludes a nut-containing nutrition bar, a cocoa-containing nutritionbar, a first olive-containing composition, a kale cracker composition, avegetable soup composition, a first vegetable broth composition, a teacomposition that includes spearmint, a energy drink composition, amicronutritional composition, and a algal oil composition.Characteristically, the daily meal portions are packaged into mealservings or into a total daily serving to be divided into meals. Thisdiet package also includes daily meal portions for a predeterminednumber of days as set forth above with the daily meal portions beingpackaged into meal servings or into a total daily serving to be dividedinto meals. In a refinement, the fasting mimicking diet package furtherincludes a mushroom soup composition, a tomato soup composition, aquinoa-containing minestrone soup composition, and a pumpkin soupcomposition. In a further refinement, the fasting mimicking diet packagefurther includes a second olive-containing composition, a secondvegetable broth composition, a bean-containing minestrone soupcomposition, a tea composition that includes spearmint and lemon, and atea composition that includes hibiscus.

As set forth above, the fasting mimicking diet packages includesspecific meal components. Typically, compositions are as follows. Thenut-containing nutrition bar includes almond meal and macadamia nuts.The cocoa-containing nutrition bar includes almond butter, almonds, andbrown rice crispy (e.g., brown puffed rice). The mushroom soupcomposition includes brown rice powder, carrots, inulin, and mushrooms.The bean-containing minestrone soup composition includes white beans,cabbage, and potatoes. The first vegetable broth composition includescarrots, maltodextrin, celery, spinach, and tomatoes. The secondvegetable broth composition includes carrots, maltodextrin, celery,spinach, soy lecithin, and tomatoes. The energy drink compositionincludes glycerin and water. The algal oil composition includesschizocatrium algae oil. The micronutrient composition includes beetroot powder, calcium carbonate, carrots, collard leaf, kale leaf, andtomatoes. In a refinement, the micronutrient composition includes Vit A,Vit C, Ca, Fe, Vit D3, Vit E, Vit K, Vit B1, Vit B2, Vit B3, Vit B5, VitB6, Vit B7, Vit B9, Vit B12, Cr, Cu, I, Mg, Mn, Mo, Se, and Zn.

In a refinement, the nut-containing nutrition bar (L-Bar Nut based)includes almond meal and macadamia nuts. In a refinement, thenut-containing nutrition bar (L-Bar Nut based) includes almond mealpreferably in an amount of 20 to 35 weight %; coconut preferably in anamount of 2 to 10 weight %; coconut oil preferably in an amount of 1 to8 weight %; flax seed meal preferably in an amount of 1 to 8 weight %;honey preferably in an amount of 10 to 30 weight %; macadamia nutspreferably in an amount of 10 to 30 weight %; pecans preferably in anamount of 10 to 25 weight %; salt preferably in an amount of 0.1 to 0.8weight %; and optionally vanilla preferably in an amount of 0.3 to 1.5weight %.

In a refinement, the cocoa-containing nutrition bar (L-Bar ChocoCrisp)includes almond butter, almonds, and brown rice crispy (PGP10235). In arefinement, the cocoa-containing nutrition bar (L-Bar ChocoCrisp)includes almond butter preferably in an amount of 10 to 25 weight %;almonds preferably in an amount of 3 to 12 weight %; brown rice crispy(PGP10235) preferably in an amount of 10 to 25 weight %; brown ricesyrup preferably in an amount of 2 to 8 weight %; chocolate liquorpreferably in an amount of 1 to 4 weight %, cocoa butter preferably inan amount of 0.4 to 1.6 weight %; cocoa powder preferably in an amountof 4 to 12 weight %; fiber syrup SF75 preferably in an amount of 18 to38 weight %, flax seed oil preferably in an amount of 1 to 3 weight %;salt preferably in an amount of 0.1 to 0.4 weight % and sugar preferablyin an amount of 1 to 6 weight %.

In a refinement, the first olive-containing composition (sea saltversion) incudes olives, olive oil, and sea salt. In a refinement, thefirst olive-containing composition (sea salt) includes lactic acidpreferably in an amount of 0.3 to 1 weight %; oil (olive) preferably inan amount of 2 to 6 weight %; olives (raw, green pitted) preferably inan amount of 50 to 97 weight %; salt (reg., kosher, sea salt) preferablyin an amount of 0.8 to 3 weight %; and thyme preferably in an amount of0.1 to 0.5 weight %.

In a refinement, the second olive-containing composition (garlicversion) incudes olives, olive oil, and garlic. In a refinement, thesecond olive-containing composition (garlic) includes garlic preferablyin an amount of 0.1 to 0.6 weight %; lactic acid preferably in an amountof 0.3 to 1 weight %; oil (olive) preferably in an amount of 2 to 6weight %; olives (raw, green pitted) preferably in an amount of 50 to 97weight %; salt (reg., kosher, sea salt) preferably in an amount of 0.8to 3 weight %; thyme preferably in an amount of 0.1 to 0.5 weight %.

In a refinement, the kale cracker composition includes kale, almonds,tapioca flour, and optionally sesame seeds. In another refinement, thekale cracker composition includes almonds preferably in an amount of 15to 40 weight %; black pepper preferably in an amount of 0.1 to 0.4weight %; chia seeds preferably in an amount of 3 to 10 weight %; chilipepper preferably in an amount of 0.4 to 1.2 weight %; cumin seedspreferably in an amount of 0.3 to 0.9 weight %; flax seeds preferably inan amount of 3 to 10 weight %; garlic preferably in an amount of 0.02 to0.04 weight %; kale preferably in an amount of 2 to 6 weight %; oil (sunflower) preferably in an about of 2 to 7 weight %; onion (powder,minced) typically in an amount of 0.3 to 0.9 weight %; oreganopreferably in an amount of 0.01 to 0.06 weight %; salt preferably in anamount of 1 to 4 weight %; sesame seeds preferably in an amount of 15 to35 weight %; sugar (coconut) preferably in an amount of 1 to 5 weight %;tapioca flour preferably in an amount of 10 to 30 weight %; vinegar(coconut) preferably in an amount of 1 to 4 weight %; water (purified)preferably in an amount of 2 to 12 weight %; and yeast extractpreferably in an amount of 0.3 to 1 weight %.

In another refinement, the kale cracker composition includes kale, flaxseeds golden, sesame seeds, and sunflower seeds. In another refinement,the apple cider vinegar preferably in an amount 1 to 3 weight %; blackpepper preferably in an amount of 0.4 to 1.3 weight %; cashewspreferably in an amount of 4 to 13 weight %; dill weed preferably in anamount of 0.4 to 1.3 weight %; flax seeds golden preferably in an amountof 13 to 40 weight %; hemp seeds preferably in an amount of 0.7 to 2weight %; kale preferably in an amount of 14 to 42 weight %; onion,white, dried, (powder, minced) preferably in an amount of 0.5 to 1.6weight %; pumpkin seeds preferably in an amount of 0.7 to 2 weight %;salt (reg. , kosher, sea salt) preferably in an amount of 0.7 to 2weight %; Sesame seeds preferably in an amount of 2 to 8 weight %;sunflower seeds preferably in an amount of 10 to 30 weight %; and yeastextract preferably in an amount of 1 to 5 weight %.

In a refinement, the vegetable soup composition includes onions,tomatoes, spinach, green tree extract, optionally rice flour, optionallybrown rice powder, optionally carrots, and optionally inulin, leeks, Ina refinement, the vegetable soup composition includes basil (whole leaf,dried) preferably in an amount of 0.3 to 0.9 weight %; brown rice powder(whole grain) preferably in an amount of 3 to 12 weight %; carrot(dehydrated, puffed, powder, pieces) preferably in an amount of 4 to 14weight %; green tea extract preferably in an amount of 0.02 to 0.06weight %; inulin preferably in an amount of 5 to 15 weight %; leeks(granules −10+40) preferably in an amount of 1 to 5 weight %; oil(olive) preferably in an amount of 1 to 6 weight %; onion (powder,minced) preferably in an amount of 4 to 15 weight %; parsley preferablyin an amount of 0.3 to 0.8 weight %; red bell peppers preferably in anamount of 1 to 5 weight %; rice flour preferably in an amount of 18 to50 weight %; salt preferably in an amount of 2 to 7 weight %; spinach(leaf, powder) preferably in an amount of 0.4 to 1.5 weight %; tomatoes(fruit powder, sun dried, granules) preferably in an amount of 4 to 14weight %; yeast extract preferably in an amount of 0.5 to 1.8 weight %.In the vegetable soup composition and any of the compositions set forthherein having rice flour, the rice flour can be glutinous ornon-glutinous, milled or unmilled.

In another refinement, the vegetable soup composition includes carrots,inulin, leeks, onions and rice flour. In a refinement, the vegetablesoup composition includes basil, whole leaf, dried preferably in anamount of 0.3 to 1 weight %; carrot (dehydrated, puffed, powder, pieces)preferably in an amount of 4 to 12 weight %; inulin preferably in anamount of 6 to 18 weight %; leeks in an amount of 1 to 5 weight %; oil(olive) preferably in an amount of 1 to 3 weight %; Onion, white, dried,(powder, minced) preferably in an amount of 10 to 30 weight %; parsleypreferably in an amount of 0.3 to 1 weight %; potato preferably in anamount of 1 to 5 weight %; red pepper preferably in an amount of 1 to 6weight %; rice flour in an amount of 13 to 40 weight %; salt (reg. ,kosher, sea salt) in an amount of 4 to 12 weight %; spinach (leaf,powder) preferably in an amount of 0.2 to 1 weight %; and tomatoes,(fruit powder, sun dried granules) preferably in an amount of 3 to 13weight %.

In a refinement, the mushroom soup composition includes mushrooms, greentea extract, optionally brown rice powder, optionally carrots, andoptionally inulin. In a refinement, the mushroom soup compositionincludes brown rice powder (whole grain) preferably in an amount of 10to 30 weight %; carrot (dehydrated, puffed, powder, pieces) preferablyin an amount of 3 to 12 weight %; green tea extract preferably in anamount of 0.02 to 0.06 weight %; inulin preferably in an amount of 3 to12 weight %; mushrooms (European mix, powder, pieces) preferably in anamount of 6 to 18 weight %; oil (olive) preferably in an amount of 1 to6 weight %; onion preferably in an amount of powder, minced) preferablyin an amount of 3 to 12 weight %; parsley preferably in an amount of 0.1to 0.5 weight %; rice flour preferably in an amount of 18 to 50 weight%; salt preferably in an amount of 2 to 8 weight %; yeast extractpreferably in an amount of 0.5to 1.5 weight %.

In another refinement, the mushroom soup composition includes carrots,inulin, mushrooms, onions, and rice flour. In another refinement, themushroom soup composition includes carrot (dehydrated, puffed, powder,pieces) preferably in an amount of 7 to 22 weight %; inulin preferablyin an amount of 7 to 22 weight %; mushrooms (European mix), (powder &pieces) dehydrated preferably in an amount of 7 to 22 weight %; oil(olive) preferably in an amount of 0.6 to 2 weight %; Onion, white,dried, (powder, minced) preferably in an amount of 7 to 22 weight %;parsley preferably in an amount of 0.3 to 0.9 weight %; potatopreferably in an amount of 0.6 to 2 weight %; rice flour preferably inan amount of 15 to 45weight %; salt (reg. , kosher, sea salt) preferablyin an amount of 6 to 18 weight %; and yeast extract preferably in anamount of 0.7 to 2.2 weight %.

In a refinement, the tomato soup composition includes tomatoes, greentea extract, optionally inulin, and optionally onions. In a refinement,the tomato soup composition (new) includes basil (whole leaf, dried)preferably in an amount of 0.2 to 0.7 weight %; brown rice powder (wholegrain) preferably in an amount of 1 to 5 weight %; green tea extractpreferably in an amount of 0.02 to 0.06 weight %; inulin preferably inan amount of 7 to 20 weight %; oil (olive) preferably in an amount of 3to 9 weight %; onion preferably (powder, minced) preferably in an amountof 4 to 12 weight %; parsley preferably in an amount of 0.1 to 0.6weight %; rice flour preferably in an amount of 18 to 50 weight %; saltpreferably in an amount of 2 to 9 weight %; tomatoes (fruit powder, sundried, granules) preferably in an amount of 12 to 36 weight %; and yeastextract preferably in an amount of 0.5 to 3 weight %.

In another refinement, the tomato soup composition includes tomatoes,inulin, olives, onions, potatoes, and rice flour. In still anotherrefinement, the tomato soup composition includes basil, whole leaf,dried preferably in an amount of 0.3 to 1 weight %; inulin preferably inan amount of 6 to 18 weight %; oil (olive) preferably in an amount of 4to 14 weight %; onion, white, dried, (powder, minced) preferably in anamount of 8 to 24 weight %; parsley preferably in an amount of 0.3 to0.9 weight %; potato preferably in an amount of 6 to 18 weight %; riceflour preferably in an amount of 9 to 27 weight %; salt (reg., kosher,sea salt) preferably in an amount of 4 to 14 weight %; tomatoes, (fruitpowder, sun dried granules) preferably in an amount of 8 to 24 weight %;and yeast extract preferably in an amount of 0.7 to 2.2 weight %.

In a refinement, the quinoa-containing minestrone soup compositionincludes quinoa, green tea extract, optionally olive oil, optionallycabbage, optionally potatoes, optionally rice flour, and optionallytomatoes and optionally no tumeric. In a refinement, thequinoa-containing minestrone soup composition includes basil (wholeleaf, dried preferably in an amount of 0.7 to 2 weight %; broccolipowder preferably in an amount of 0.6 to 2 weight %; cabbage white(flakes) preferably in an amount of 3 to 10 weight %; carrot(dehydrated, puffed, powder, pieces) preferably in an amount of 3 to 10weight %; celery preferably in an amount of 1 to 4 weight %; celeryseeds (powder) preferably in an amount of 0.07 to 0.2 weight %; garlicpreferably in an amount of 0.7 to 2 weight %; green tea extractpreferably in an amount of 0.02 to 0.06 weight %; inulin preferably inan amount of 1 to 5 weight % ; leeks (granules −10+40), preferably in anamount of 0.7 to 2 weight %; oil (olive) preferably in an amount of 0.6to 2 weight %; onion (powder, minced) preferably in an amount of 2 to 8weight %; peas preferably in an amount of 3 to 10 weight %; potatopreferably in an amount of 7 to 20 weight %; quinoa preferably in anamount of 7 to 20 weight %; rice flour preferably in an amount of 7 to20 weight %; salt, preferably in an amount of 1 to 6 weight %; spinach(leaf, powder) preferably in an amount of 0.5 to 2 weight %; tomatoes(fruit powder, sun dried, granules) preferably in an amount of 2 to 6weight %; yeast extract preferably in an amount of 0.6 to 2 weight %;zucchini (powder, diced) preferably in an amount of 2 to 8 weight %.

In another refinement, the quinoa-containing minestrone soup includesquinoa, cabbage, potatoes, and rice flour. In still another refinement,the quinoa-containing minestrone soup includes basil, whole leaf, driedpreferably in an amount of 0.7 to 2.2 weight %; broccoli powderpreferably in an amount of 0.7 to 2.2 weight %; cabbage white (flakes)preferably in an amount of 0.6 to 2.2 weight %; carrot (dehydrated,puffed, powder, pieces) preferably in an amount of 3 to 10 weight %;celeriac preferably in an amount of 2 to 6 weight %; celery seeds powderpreferably in an amount of 0.6 to 1.8 weight %; garlic preferably in anamount of 1 to 3 weight %; Onion, white, dried, (powder, minced)preferably in an amount of 3 to 9 weight %; peas preferably in an amountof 3 to 10 weight %; potato preferably in an amount of 6 to 20 weight %;quinoa preferably in an amount of 8 to 23 weight %; rice flourpreferably in an amount of 7 to 22 weight %; salt (reg., kosher, seasalt) preferably in an amount of 2 to 7 weight %; savoy cabbagepreferably in an amount of 3 to 10 weight %; spinach (leaf, powder)preferably in an amount of 0.7 to 2.2 weight %; turmeric preferably inan amount of 0.6 to 1.8 weight %; yeast extract preferably in an amountof 3 to 10 weight %; and zucchini (powder, diced) preferably in anamount of 1 to 5 weight %.

In a refinement, the bean-containing minestrone soup compositionincludes white beans (e.g., great northern beans), great tea extract,optionally cabbage, and optionally potatoes. In a refinement, thebean-containing minestrone soup composition includes beans (greatnorthern) preferably in an amount of 3 to 10 weight %; cabbage white(flakes) preferably in an amount of 2 to 8 weight %; carrot (dehydrated,puffed, powder, pieces) preferably in an amount of 2 to 8 weight %;celery preferably in an amount of 1 to 4 weight %; green tea extractpreferably in an amount of 0.02 to 0.06 weight %; inulin preferably inan amount of 2 to 10 weight %; leeks (granules −10+40) preferably in anamount of 2 to 7 weight %; oil (olive) preferably in an amount of 2 to 7weight %; onion (powder, minced) preferably in an amount of 2 to 7weight %; parsley preferably in an amount of 0.2 to 1 weight %; peaspreferably in an amount of 3 to 9 weight %; potato preferably in anamount of 15 to 45 weight %; rice flour preferably in an amount of 6 to18 weight %; salt preferably in an amount of 2 to 8 weight %; spinach(leaf, powder) preferably in an amount of 0.5 to 1.5 weight %; tomatoes(fruit powder, sun dried, granules) preferably in an amount of 2 to 7weight %; and yeast extract preferably in an amount of 0.5 to 1.5 weight%.

In a refinement, the bean-containing minestrone soup compositionincludes brown beans, carrots, peas, potato, and rice flour. In anotherrefinement, the bean-containing minestrone soup composition includescarrot (dehydrated, puffed, powder, pieces) preferably in an amount of 4to 14 weight %; celeriac preferably in an amount of 1 to 5 weight %;celery preferably in an amount of 0.5 to 1.6 weight %; leeks preferablyin an amount of 2 to 8 weight %; oil (olive) preferably in an amount of2 to 8 weight %; Onion, white, dried, (powder, minced) preferably in anamount of 3 to 10 weight %; parsley preferably in an amount of 0.5 to1.5 weight %; peas preferably in an amount of 5 to 18 weight %; potatopreferably in an amount of 8 to 24 weight %; rice flour preferably in anamount of 5 to 18 weight %; salt (reg., kosher, sea salt) preferably inan amount of 4 to 14 weight %; spinach (leaf, powder) preferably in anamount of 0.5 to 1.5 weight %; tomatoes, (fruit powder, sun driedgranules) preferably in an amount of 0.9 to 2.8 weight %; turmericpreferably in an amount of 0.3 to 1.2 weight %; and yeast extractpreferably in an amount of 0.5 to 1.5 weight %.

In a refinement, the pumpkin soup composition includes pumpkin, greentree extract, optionally rice flour, optionally carrots, and optionallybrown rice powder. In a refinement, the pumpkin soup compositionincludes (new) includes brown rice powder (whole grain) preferably in anamount of 3 to 9 weight %; carrot (dehydrated, puffed, powder, pieces)preferably in an amount of 2 to 8 weight %; green tea extract preferablyin an amount of 0.02 to 0.06 weight %; inulin preferably in an amount of2 to 10 weight %; oil (olive) preferably in an amount of 1 to 7 weight%; onion (powder, minced) preferably in an amount of 1.0 to 3 weight %;pumpkin powder preferably in an amount of 20 to 60 weight %; rice flourpreferably in an amount of 15 to 45 weight %; salt preferably in anamount of 2 to 10 weight %; and yeast extract preferably in an amount of0.3 to 1 weight %.

In a refinement, the first vegetable broth includes carrots,maltodextrin, celery, spinach, and tomatoes. In a refinement, the firstvegetable broth includes carrot (dehydrated, puffed, powder, pieces)preferably in an amount of 6 to 18 weight %; celery preferably in anamount of 3 to 10 weight %; garlic preferably in an amount of 3 to 10weight %; maltodextrin preferably in an amount of 8 to 25 weight %; oil(canola) preferably in an amount of 0.5 to 2 weight %; onion (powder,minced) preferably in an amount of 6 to 18 weight %; parsley preferablyin an amount of 3 to 10 weight %; potato preferably in an amount of 1 to3 weight %; salt preferably in an amount of 7 to 21 weight %; spinach(leaf, powder) preferably in an amount of 3 to 10 weight %; tomatoes(fruit powder, sun dried, granules) preferably in an amount of 6 to 18weight %; and yeast extract preferably in an amount of 1 to 6 weight %.

In a refinement, the second vegetable broth (chicken flavoring) includescarrots, chicken flavoring, maltodextrin, celery, spinach, soy lecithin,and tomatoes. In a refinement, the second vegetable broth compositionincludes carrot (dehydrated, puffed, powder, pieces) preferably in anamount of 3 to 10 weight %; celery preferably in an amount of 3 to 12weight %; garlic preferably in an amount of 3 to 9 weight %;maltodextrin preferably in an amount of 8 to 25 weight %; oil (canola)preferably in an amount of 0.5 to 2 weight %; onion preferably in anamount of powder, minced) preferably in an amount of 3 to 12 weight %;parsley preferably in an amount of 3 to 10 weight %; potato preferablyin an amount of 1 to 6 weight %; salt preferably in an amount of 8 to 25weight %; soy lecithin preferably in an amount of 0.5 to 3 weight %;spinach (leaf, powder) preferably in an amount of 3 to 12 weight %;tomatoes (fruit powder, sun dried, granules) preferably in an amount of6 to 18 weight %; xanthan gum preferably in an amount of 0.5 to 4 weight%; and yeast extract preferably in an amount of 4 to 12 weight %.

In a refinement, the energy drink composition includes glycerinpreferably in an amount of 20 to 60 weight %; water (purified)preferably in an amount of 40 to 80 weight %.

In a refinement, the tea composition that includes spearmint includesspearmint leaves organic preferably in an amount of 70 to 100 weight %.

In a refinement, the tea composition that includes lemon and spearmintincludes lemon myrtle organic preferably in an amount of 3 to 12 weight%; lemon peel organic preferably in an amount of 10 to 25 weight %;spearmint leaves organic preferably in an amount of 50 to 95 weight %.

In a refinement, the tea composition that includes hibiscus includeshibiscus tea leaves organic preferably in an amount of 80 to 100 weight%.

In a refinement, the algal oil composition includes schizocatrium algaeoil (DHA Omega-3) preferably in an amount of 80 to 100 weight %.

In a refinement, the nutrient replenishment composition (NR-1) includesbeet root powder, calcium carbonate, carrots, collard leaf, kale leaf,and tomatoes. In a refinement, the nutrient replenishment composition(NR-1) includes ascorbic acid preferably in an amount of 1 to 3 weight%; beet root powder preferably in an amount of 6 to 20 weight %; betacarotene preferably in an amount of 0.05 to 0.15 weight %; calciumcarbonate preferably in an amount of 6 to 20 weight %; carrot(dehydrated, puffed, powder, pieces) preferably in an amount of 6 to 20weight %; cholecaliciferol preferably in an amount of 0.00 weight %;chromuim Picolinate preferably in an amount of 0.00 weight %; collardleaf powder preferably in an amount of 6 to 20 weight %; cupric sulfatepreferably in an amount of 0.01 to 0.06 weight %; cyanocobalamin, 0.00;D1-alpha tocopherol acetate preferably in an amount of 0.3 to 1 weight%; ferrous fumarate preferably in an amount of 0.2 to 1 weight %; folicacid preferably in an amount of 0.00 weight %; kale leaf preferably inan amount of 6 to 20 weight %; magnesium stearate preferably in anamount of 1 to 6 weight %; manganese sulfate preferably in an amount of0.04 to 0.08 weight %; niacinamide preferably in an amount of 0.3 to 1weight %; pantothenic acid preferably in an amount of 0.1 to 0.6 weight%; phytonadione preferably in an amount of 0.00 weight %; potassiumiodine preferably in an amount of 0 weight %; pyriodoxine HCl preferablyin an amount of 0.03 to 0.1 weight %; riboflavin preferably in an amountof 0.02 to 0.1 weight %; sodium molybdate preferably in an amount of0.00 weight %; sodium selenate preferably in an amount of 0.00 weight %;spinach (leaf, powder) preferably in an amount of 6 to 20 weight %;thiamine mononitrate preferably in an amount of 0.02 to 0.1 weight %;tomatoes (fruit powder, sun dried, granules) preferably in an amount of6 to 20 weight %; tribasic calcium phosphate preferably in an amount of0.5 to 2 weight %; and zinc oxide preferably in an amount of 0.2 to 0.8weight %.

In a variation, the each of the components of the fasting mimicking dietpackage and therefore the fasting mimicking diet, is substantiallygluten free (e.g., each component has less than 20 ppm gluten) or verylow gluten (e.g., each component has 20-100 ppm). In other variations,each of the components are provided in a serving size from 20 to 60 g.In other variations, the nut-containing nutrition bar is provided in aserving size from 30 to 60 g; cocoa-containing nutrition bar is providedin a serving size from 15 to 40 g; the olive containing composition (seasalt version) in a serving size from 10 to 20 g; the olive containingcomposition (garlic version) in a serving size from 10 to 20 g; kalecracker composition is provides in a serving size from 30 to 60 g; Inanother variation, the kale cracker compositions are provided in aserving size from 20 to 50 g; the vegetable soup compositions areprovided in a serving size from 20 to 50 g; the mushroom soupcompositions are provided in a serving size from 20 to 50 g; the tomatosoup compositions are provided in a serving size from 20 to 50 g; thebean-containing minestrone soup compositions are provided in a servingsize from 20 to 50 g; the quinoa-containing minestrone soup compositionsare provided in a serving size from 20 to 50 g; the pumpkin soupcompositions are provided in a serving size from 20 to 50; the firstvegetable both compositions are provided in a serving size from 5 to 15;the second vegetable both compositions are provided in a serving sizefrom 3 to 15; and Energy Drink composition is provided in serving sizeof 1 to 5 oz.

The table set forth below provides a schedule of administration for twoFMD meal plans to be administered to a subject. The Prolon Meal plan isuseful for weight loss, treating or preventing hypertension, metabolicdisease, diabetes, and the like. The Chemolieve meal plan is useful foralleviating the side effect of chemotherapy. Therefore, the dietpackages set forth herein can include instruction providing theschedules and instructions for administering the FMD to treat variousconditions as set forth in the methods below.

Table of Meal Schedules MEAL PLAN - PROLON US MEAL PLAN CHEMOLIEVE USDAY DAY DAY DAY DAY DAY DAY DAY DAY DAY COMPONENTS (single servings) 1 23 4 5 1 2 3 4 5 nut-containing nutrition bar (L-Bar 2 1 1 1 1 1 — — — 1Nut based) cocoa-containing nutrition bar (L- 1 1 — 1 — — — — — — BarChocoCrisp) - .83 oz. First olive-containing composition 1 1 — 1 — — — —— — (Sea Salt) - 0.73 oz Second olive-containing — 1 — 1 — — — — — —composition (Garlic) - 0.73 oz kale cracker composition, (35 g) 1 — 1 —1 1 1 — — — vegetable soap composition — — — 1 — 1 — — 1 — mushroom soapcomposition — 1 — — — — 1 — — — tomato soap composition 1 — 1 — 1 1 — —— 1 quinoa-containing minestrone soup — 1 — 1 — — — — — — compositionbean-containing minestrone soup 1 — 1 — 1 — — — — — composition pumpkinsoup composition, — — — — — — — 1 — — First vegetable broth composition— — — — — — — 1 — 1 Second vegetable broth — — — — — — 1 — 1 —composition (chicken) Energy Drink — 1 1 1 1 1 1 — — — Tea - Spearmint 11 1 1 1 1 1 1 1 1 Tea - Lemon Spearmint 1 1 1 1 1 1 1 1 1 1 Tea -Hibiscus — 2 2 2 2 — — — — — Algal oil 1 — — — 2 2 — — — 1 NR-1 2 1 1 11 2 1 1 1 1

While exemplary embodiments are described above, it is not intended thatthese embodiments describe all possible forms of the invention. Rather,the words used in the specification are words of description rather thanlimitation, and it is understood that various changes may be madewithout departing from the spirit and scope of the invention.Additionally, the features of various implementing embodiments may becombined to form further embodiments of the invention.

REFERENCES

1. Cosmi, L., Liotta, F., Maggi, E., Romagnani, S. & Annunziato, F. Th17and non-classic Th1 cells in chronic inflammatory disorders: two sidesof the same coin. International archives of allergy and immunology 164,171-177, doi:10.1159/000363502 (2014).

2. Dornmair, K., Goebels, N., Weltzien, H. U., Wekerle, H. & Hohlfeld,R. T-cell-mediated autoimmunity: novel techniques to characterizeautoreactive T-cell receptors. The American journal of pathology 163,1215-1226, doi:10.1016/S0002-9440(10)63481-5 (2003).

3. Fasano, A. & Shea-Donohue, T. Mechanisms of disease: the role ofintestinal barrier function in the pathogenesis of gastrointestinalautoimmune diseases. Nature clinical practice. Gastroenterology &hepatology 2, 416-422, doi:10.1038/ncpgasthep0259 (2005).

4. Eksioglu-Demiralp, E. et al. Phenotypic characteristics of B cells inBehcet's disease: increased activity in B cell subsets. The Journal ofrheumatology 26, 826-832 (1999).

5. Holmen, N., Isaksson, S., Simren, M., Sjovall, H. & Ohman, L.CD4+CD25+regulatory T cells in irritable bowel syndrome patients.Neurogastroenterology and motility: the official journal of the EuropeanGastrointestinal Motility Society 19, 119-125,doi:10.1111/j.1365-2982.2006.00878.x (2007).

6. Xavier, R. J. & Podolsky, D. K. Unravelling the pathogenesis ofinflammatory bowel disease. Nature 448, 427-434, doi:10.1038/nature06005(2007).

7. Rovedatti, L. et al. Differential regulation of interleukin 17 andinterferon gamma production in inflammatory bowel disease. Gut 58,1629-1636, doi:10.1136/gut.2009.182170 (2009).

8. Ueno, A. et al. Increased prevalence of circulating novel IL-17secreting Foxp3 expressing CD4+T cells and defective suppressivefunction of circulating Foxp3+ regulatory cells support plasticitybetween Th17 and regulatory T cells in inflammatory bowel diseasepatients. Inflammatory bowel diseases 19, 2522-2534,doi:10.1097/MIB.0b013e3182a85709 (2013).

9. Jin, D. et al. Manipulation of Microbiome, a Promising Therapy forInflammatory Bowel Diseases. Journal of Clinical & Cellular Immunology05(04). doi:10.4172/2155-9899.1000234 (2014).

10. Mcilroy, J., Ianiro, G., Mukhopadhya, I., Hansen, R., & Hold, G. L.Review article: the gut microbiome in inflammatory bowel disease-avenuesfor microbial management. Alimentary Pharmacology & Therapeutics 47(1),26-42. doi:10.1111/apt.14384 (2017).

11. Yang, J-Y., Lee, Y-S., Kim, Y., Lee, S-H., Ryu, S. et al. (2017).Gut commensal Bacteroides acidifaciens prevents obesity and improvesinsulin sensitivity in mice. Mucosal immunology, 10, 104-116. 12.O'Neill, I., Schofield, Z., & Hall, L. (2017). Exploring the role of themicrobiota member Bifidobacterium in modulating immune-linked diseases.Emerging topics in life sciences, 1(4), 333-349. 13. Dupaul-Chicoine,J., Yeretssian, G., Doiron, K., Bergstrom, K. S., McIntire, C. R.,LeBlanc, P. M., Meunier, C., Turbide, C., Gros, P., Beauchemin, N., etal. (2010). Control of intestinal homeostasis, colitis, andcolitis-associated colorectal cancer by the inflammatory caspases.Immunity 32, 367-378.

14. Koblansky, A. A., Truax, A. D., Liu, R., Montgomery, S. A., Ding,S., Wilson, J. E., Brickey, W. J., M€uhlbauer, M., McFadden, R. M., Hu,P., et al. (2016). The innate immune receptor NLRX1 functions as a tumorsuppressor by reducing colon tumorigenesis and key tumor-promotingsignals. Cell Rep. 14, 2562-2575.

PUBLICATIONS

1. TITLE Manuscript Number: THELANCETNEUROLOGY-D-16-00304

2. Choi I Y, Piccio L, Childress P, Bollman B, Ghosh A, Brandhorst S,Suarez J, Michalsen A, Cross A H, Morgan T E, Wei M, Paul F, Bock M,Longo V D. A Diet Mimicking Fasting Promotes Regeneration and ReducesAutoimmunity and Multiple Sclerosis Symptoms. Cell reports. 2016;15(10):2136-46. doi: 10.1016/j.celrep.2016.05.009. PubMed PMID:27239035; PMCID: PMC4899145.

3. Brandhorst S, Choi I Y, Wei M, Cheng C W, Sedrakyan S, Navarrete G,Dubeau L, Yap LP, Park R, Vinciguerra M, Di Biase S, Mirzaei H, MirisolaM G, Childress P, Ji L, Groshen S, Penna F, Odetti P, Perin L, Conti PS, Ikeno Y, Kennedy B K, Cohen P, Morgan T E, Dorff T B, Longo V D. APeriodic Diet that Mimics Fasting Promotes Multi-System Regeneration,Enhanced Cognitive Performance, and Healthspan. Cell Metab. 2015;22(1):86-99. doi: 10.1016/j.cmet.2015.05.012. PubMed PMID: 26094889;PMCID: PMC4509734.

4. Levine M E, Suarez J A, Brandhorst S, Balasubramanian P, Cheng C W,Madia F, Fontana L, Mirisola M G, Guevara-Aguirre J, Wan J, Passarino G,Kennedy B K, Wei M, Cohen P, Crimmins E M, Longo V D. Low protein intakeis associated with a major reduction in IGF-1, cancer, and overallmortality in the 65 and younger but not older population. Cell Metab.2014; 19(3):407-17. doi: 10.1016/j.cmet.2014.02.006. PubMed PMID:24606898; PMCID: PMC3988204.

5. Longo V D, Mattson M P. Fasting: molecular mechanisms and clinicalapplications. Cell Metab. 2014; 19(2):181-92. doi:10.1016/j.cmet.2013.12.008. PubMed PMID: 24440038; PMCID: 3946160.

6. Wei M, Brandhorst S, Shelehchi M, Mirzaei H, Cheng C W, Budniak J,Groshen S, Mack W J, Guen E, Di Biase S, Cohen P, Morgan T E, Dorff T,Hong K, Michalsen A, Laviano A, Longo V D. Fasting-mimicking diet andmarkers/risk factors for aging, diabetes, cancer, and cardiovasculardisease. Sci Transl Med. 2017; 9(377). PubMed PMID: 28202779.

What is claimed is:
 1. A probiotic composition for gastrointestinalautoimmune and/or inflammatory disease, the probiotic compositioncomprising: a bacterial component that is a combination of Bacteroidesacidifaciens and Bifidobacterium choerinum such that the Bifidobacteriumchoerinum enhances probiotic activity of Bacteroides acidifaciens to beadministered for prevention and treatment of gastrointestinal autoimmuneand/or inflammatory disease; and a protective component that stabilizesthe bacterial component.
 2. The probiotic composition of claim 1 whereinthe bacterial component further comprises gut microbiota strainsisolated from genera selected from the group consisting ofLactobacillus, Allobaculum, Bifidobacterium, and combinations thereof.3. The probiotic composition of claim 2 wherein each bacterial componentis present in an amount from about 10³ to about 10¹⁴ CFU/g of eachbacterial component.
 4. The probiotic composition of claim 1 wherein theprotective component is selected from the group consisting of protectivecarriers, protective coating, and protective encapsulants.
 5. Theprobiotic composition of claim 1 wherein the protective component is aliquid.
 6. The probiotic composition of claim 5 wherein the liquid is ahydroxylated hydrocarbon carrier.
 7. The probiotic composition of claim5 wherein the liquid is a polyol.
 8. The probiotic composition of claim1 wherein the protective component includes a stabilization agent thatinteracts with or encapsulates the bacterial component.
 9. The probioticcomposition of claim 8 wherein the stabilization agent is selected fromthe group consisting of adonitol, betaine, carbohydrates, proteins,amino acids, mixtures of sugar and protein, gums and skim milk, andcombinations thereof.
 10. The probiotic composition of claim 9 whereinthe stabilization agent is dissolved or dispersed in a liquid.
 11. Theprobiotic composition of claim 1 wherein the protective componentencapsulates the bacterial component.
 12. The probiotic composition ofclaim 11 wherein the bacterial component is encapsulated by spraydrying, fluidized bed drying, or vacuum drying.
 13. The probioticcomposition of claim 11 wherein the protective component includes anencapsulating agent selected from the group consisting of maltodextrins,skim milk, reconstituted skim milk, casein, soybean protein, trehalose,and combinations thereof.
 14. The probiotic composition of claim 1further comprising an additive selected from the group consisting of anysuitable adjuvants, excipients, additives, additional therapeuticagents, bioavailability enhancers, side-effect suppressing components,diluents, buffers, flavoring agents, binders, preservatives or otheringredients and combinations thereof.
 15. The probiotic composition ofclaim 1 incorporated into a capsule.